Elsevier Editorial System(tm) for Crop Protection Manuscript Draft Manuscript Number: CROPRO-D-17-00239 Title: Identification of a newly described member of the tribe Erythroneurini as a potential vector of the Côte d'Ivoire lethal yellowing phytoplasma in coconut palms sole or in mixed infection with a 'Candidatus Phytoplasma asteris'-related strain. Article Type: Full Length Article Section/Category: Crop pathogens such as fungi, oomycetes, bacteria, viruses, other microbes and nematodes Keywords: coconut lethal yellowing, phytoplasma, potential vector, 16SrXXII, Côte d'Ivoire, Nedotepa Corresponding Author: Dr. Yaima Arocha Rosete, PhD Corresponding Author's Institution: First Author: Koffi Eric Kwadjo, Dr Order of Authors: Koffi Eric Kwadjo, Dr; N’Djiha Isabelle Beugré, MSc; Christopher H. Dietrich, Prof.; Adaba Tano Thierry Kodjo, MSc; Hortense Atta Diallo, Prof; Ndede Yankey, Dr.; Sylvester Dery, Prof.; Michael Wilson, Dr; Jean Louis Konan Konan, Dr; Nicoletta Contaldo, Dr; Samanta Paltrinieri, Dr; Assunta Bertaccini, Prof.; Yaima Arocha Rosete, PhD Abstract: Over 300 Hemiptera specimens were collected using sweep nets and hand-made aspirators from coconut palm fronds in six villages of Grand-Lahou. Eight families were identified including Aphrophoridae, Achilidae, Derbidae, Flatidae, Membracidae, Pentatomidae, Tropiduchidae, and Cicadellidae, the latter being the most abundant throughout the surveyed villages. PCR assays with primers targeting the 16S rRNA and the secA translocation protein genes yielded PCR amplicons from 216 out of 296 (73%) of the tested specimens of a newly identified cicadellid leafhopper, Nedotepa curta Dmitriev. PCR amplicons were purified, cloned and sequenced. The 16S rDNA and secA sequences from N. curta showed a 99% sequence identity with those of the phytoplasma identified in coconut- growing villages of Grand-Lahou, which suggested N. curta as a potential vector for the CILY phytoplasma. Phytoplasmas of group 16SrI 'Candidatus Phytoplasma asteris'-related were identified from phytoplasma-infected coconut palms infected by the Côte d'Ivoire lethal yellowing phytoplasma and N. curta specimens from Badadon and Yaokro villages, as well as from the weeds Dalbergia saxatilis and Baphia nitida from Badadon. Results indicate that mixed infection of both 16SrXXII-B and 16SrI phytoplasmas is occurring in coconut palms affected by CILY in Grand-Lahou, which may impact disease management and control. Suggested Reviewers: Nicola Mori Prof. University of Padova [email protected] Phytoplasma Entomologist Norma Elena Leyva Lopez Prof. CIDIIR, Mexico [email protected] Specialized on phytoplasma disease epidemiology, diagnosis and control Rosa La Rosa Prof. University of Catania [email protected] Specialized on phytoplasma epidemiology and diagnosis Helena Montano Dr University of Rio de Janeiro [email protected] Specialized on phytoplasma, epidemiology, characterization and diagnosis Cover Letter Crop Protection Cover Letter This is a research article entitled ‘Identification of a newly described genus and species of the tribe Erythroneurini as a potential vector of the Côte d’Ivoire lethal yellowing phytoplasma in coconut palms sole or in mixed infection with a ‘Candidatus Phytoplasma asteris’-related strain’. This is the first time that we publish in Crop Protection. The research described in this paper is novel and has not been published elsewhere nor submitted to any other journal for consideration of publication. Authors declare no conflict of interest. The list of authors is as follows: 1. Dr. Koffi Eric Kwadjo, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d'Ivoire, [email protected] 2. Ms. N’Djiha Isabelle Beugré, Station de Recherche “Marc Delorme”, Centre National de Recherche Agronomique (CNRA), 07 BP 13, Abidjan, Côte d'Ivoire, [email protected] 3. Dr. Christopher H. Dietrich, Illinois Natural History Survey, University of Illinois 1816 S Oak St., Champaign, IL 61820 USA, [email protected] 4. Mr. Adaba Tano Thierry Kodjo, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d'Ivoire, [email protected] 5. Prof. Hortense Atta Diallo, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d'Ivoire, [email protected] 6. Dr. Ndede Yankey, 4Council for Scientific Research Program–Oil Palm Research Institute– Coconut Research Program (CSIR-OPRI), Off Well Road Box 254, Sekondi, Western Region, Ghana, [email protected] 7. Dr. Sylvester Dery, 4Council for Scientific Research Program–Oil Palm Research Institute– Coconut Research Program (CSIR-OPRI), Off Well Road Box 254, Sekondi, Western Region, Ghana, [email protected] 8. Dr. Michael Wilson, Department of Biodiversity and Systematic Biology, Museum of Wales, Cardiff, CF10 3NP, United Kingdom, [email protected] 9. Prof. Jean Louis Konan Konan, Station de Recherche “Marc Delorme”, Centre National de Recherche Agronomique (CNRA), 07 BP 13, Abidjan, Côte d'Ivoire, [email protected] 10. Dr. Nicoletta Contaldo, Department of Agricultural Sciences, Alma Mater Studiorum - University of Bologna viale Fanin 42, 40127 Bologna, Italy, [email protected] 11. Dr. Samanta Paltrinieri, Department of Agricultural Sciences, Alma Mater Studiorum - University of Bologna viale Fanin 42, 40127 Bologna, Italy, [email protected] 12. Prof. Assunta Bertaccini, Department of Agricultural Sciences, Alma Mater Studiorum - University of Bologna viale Fanin 42, 40127 Bologna, Italy, [email protected] 13. Dr. Yaima Arocha Rosete, 7Sporometrics, 219 Dufferin Street, Suite 20C, Toronto, ON M6K 3J1, Canada. Corresponding author: [email protected], Tel: 416-516-1660. *Manuscript Click here to view linked References 1 Identification of a newly described member of the tribe Erythroneurini as a potential 2 vector of the Côte d’Ivoire lethal yellowing phytoplasma in coconut palms sole or in mixed 3 infection with a ‘Candidatus Phytoplasma asteris’-related strain. 4 5 Koffi Eric Kwadjoa, N’Djiha Isabelle Beugréb, Christopher H. Dietrichc, Adaba Tano 6 Thierry Kodjob, Hortense Atta Diallob, Ndede Yankeyd, Sylvester Deryd, Michael Wilsone, 7 Jean Louis Konan Konanc, Nicoletta Contaldof, Samanta Paltrinierif, Assunta Bertaccinif, 8 and Yaima Arocha Roseteg* 9 10 aUniversité Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d'Ivoire 11 bStation de Recherche “Marc Delorme”, Centre National de Recherche Agronomique (CNRA), 12 07 BP 13, Abidjan, Côte d'Ivoire 13 cIllinois Natural History Survey, University of Illinois 1816 S Oak St., Champaign, IL 61820 14 USA 15 dCouncil for Scientific Research Program–Oil Palm Research Institute–Coconut Research 16 Program (CSIR-OPRI), Off Well Road Box 254, Sekondi, Western Region, Ghana 17 eDepartment of Biodiversity and Systematic Biology, Museum of Wales, Cardiff, CF10 3NP, 18 United Kingdom, 19 fDepartment of Agricultural Sciences, Alma Mater Studiorum - University of Bologna viale 20 Fanin 42, 40127 Bologna, Italy 21 gSporometrics, 219 Dufferin Street, Suite 20C, Toronto, ON M6K 3J1, Canada 22 *Email: [email protected], Tel: 416-516-1660. 23 24 Abstract 25 Over 300 Hemiptera specimens were collected using sweep nets and hand-made aspirators from 26 coconut palm fronds in six villages of Grand-Lahou. Eight families were identified including 27 Aphrophoridae, Achilidae, Derbidae, Flatidae, Membracidae, Pentatomidae, Tropiduchidae, and 28 Cicadellidae, the latter being the most abundant throughout the surveyed villages. PCR assays 29 with primers targeting the 16S rRNA and the secA translocation protein genes yielded PCR 30 amplicons from 216 out of 296 (73%) of the tested specimens of a newly identified cicadellid 31 leafhopper, Nedotepa curta Dmitriev. PCR amplicons were purified, cloned and sequenced. The 32 16S rDNA and secA sequences from N. curta showed a 99% sequence identity with those of the 33 phytoplasma identified in coconut-growing villages of Grand-Lahou, which suggested N. curta 34 as a potential vector for the CILY phytoplasma. Phytoplasmas of group 16SrI ‘Candidatus 35 Phytoplasma asteris’-related were identified from phytoplasma-infected coconut palms infected 36 by the Côte d’Ivoire lethal yellowing phytoplasma and N. curta specimens from Badadon and 37 Yaokro villages, as well as from the weeds Dalbergia saxatilis and Baphia nitida from Badadon. 38 Results indicate that mixed infection of both 16SrXXII-B and 16SrI phytoplasmas is occurring in 39 coconut palms affected by CILY in Grand-Lahou, which may impact disease management and 40 control. 41 Keywords: coconut lethal yellowing, phytoplasma, potential vector, 16SrXXII, Côte d’Ivoire, 42 Nedotepa 43 44 1. Introduction 45 Côte d’Ivoire lethal yellowing (CILY) of coconut palm was first reported in 2013 in Grand- 46 Lahou and since then it has rapidly spread to several coconut-growing villages where over 400 47 ha have been destroyed and another 7,000 ha are under threat (Arocha-Rosete et al., 2014). 48 Lethal yellowing (LY)-like diseases of palms have been associated with a number of 49 phytoplasmas (Sullivan and Harrison, 2013) worldwide that have killed millions of palms in the 50 last 40 years. 51 Phytoplasmas are bacteria of the class Mollicutes transmitted by phloem-feeding insect 52 species within the order Hemiptera, particularly Cicadellidae (leafhoppers), but also Cercopidae, 53 Cixiidae, Derbidae, Delphacidae, and Psyllidae (Weintraub and Beanland, 2006). Phytoplasma 54 transmission by hemipteran vectors has previously been shown to be persistent and propagative, 55